Table 2. Residual activity and auto-inactivation of GAD chimaeras.
Residual activity±S.D. was calculated by dividing the glutamate turnover of the 10 min auto-inactivated sample by the untreated sample. All assays were performed at least four times. Auto-activation was calculated as 100%-% residual activity. Increase or decrease was calcualted as the fold difference in percentage auto-inactivation compared with respective WT (wild-type) calculated from the percentage residual activity.
| GAD chimaera | Residual activity (%) | Auto-inactivation (%) | Fold increase (↑) or decrease (↓) in auto-inactivation§ |
|---|---|---|---|
| GAD67WT | 101.8±2.4 | 0±2.4 | N/A |
| GAD67Tyr292Phe_65loop* | 84.3±4.8 | 15.7±4.8 | ↑ 1.21 |
| GAD67Tyr292Phe_65loop_Val444Leu | 51.1±6.2 | 48.9±6.2 | ↑ 1.99 |
| GAD6765CT† | 80.1±4.3 | 19.9±4.3 | ↑ 1.27 |
| GAD67Tyr292Phe_65loop_Val444Leu_65CT | 41.9±1.9 | 58.1±1.9 | ↑ 2.43 |
| GAD65WT | 45.2±3.6 | 54.8±3.6 | NA |
| GAD65Phe283Tyr_67loop‡ | 79.5±5.5 | 20.5±5.5 | ↓ 1.76 |
| GAD65Phe283Tyr_67loop_Leu435Val | 76.9±2.7 | 23.1±2.7 | ↓ 1.70 |
| GAD6567CT§ | 95.7±10.3 | 4.3±10.3 | ↓ 2.12 |
| GAD65Phe283Tyr_67loop_Leu435Val_67CT | 99.8±8.0 | 0.2±8.0 | ↓ 2.21 |
*65loop=GAD65 residues 422–433.
†65CT=GAD65 residues 454–585.
‡67loop=GAD67 residues 431–442.
§67CT=GAD67 residues 463–594.